Super-Elastic Carbonized Mushroom Aerogel for Management of Uncontrolled Hemorrhage

Abstract

Uncontrolled hemorrhage is still the most common cause of potentially preventable death after trauma in prehospital settings. However, there rarely are hemostatic materials that can achieve safely and efficiently rapid hemostasis simultaneously. Here, new carbonized cellulose-based aerogel hemostatic material is developed for the management of noncompressible torso hemorrhage, the most intractable issue of uncontrolled hemorrhage. The carbonized cellulose aerogel is derived from the Agaricus bisporus after a series of processing, including cutting, carbonization, purification, and freeze-drying. In vitro, the carbonized cellulose aerogels with porous structure show improved hydrophilicity, good blood absorption, and coagulation ability, rapid shape recoverable ability under wet conditions. And in vivo, the carbonized aerogels show effective hemostatic ability in both small and big animal serious hemorrhage models. The amount of blood loss and the hemostatic time of carbonized aerogels are all better than the positive control group. Moreover, the mechanism studies reveal that the good hemostatic ability of the carbonized cellulose aerogel is associated with high hemoglobin binding efficiency, red blood cell absorption, and platelets absorption and activation. Together, the carbonized aerogel developed in this study could be promising for the management of uncontrolled hemorrhage.

Keywords: cellulose; hemostatic materials; platelet activation; porous structure; uncontrolled hemorrhage.

Scheme 1

The schematic diagram is illustrating the preparation of carbonized mushroom aerogel and its application on hemostasis. A) The flow chart detailed introduces the manufacturing processes of carbonized mushroom aerogel, including cutting, carbonization, purification, freeze drying, and grinding. B) The carbonized mushroom aerogel powders are appropriate for the hemostasis of superficial wounds. C) The super‐elastic carbonized mushroom aerogel is suitable for the hemostasis of noncompressible torso hemorrhage.

Figure 1

Physical characterization of carbonized mushroom aerogels. A) SEM images of the internal structure of carbonized mushroom aerogels that generated under 160 °C, 180 °C, 200 °C for 8, 10, and 12 h respectively. B) The average pore size of different carbonized mushroom aerogels. C) The porosity of the different carbonized mushroom aerogels and positive controls, including gelatin, Gauze, and charcoal. D) The EDX elemental analysis of the Agaricus bisporus before and after carbonization. E) The quantification of major elemental content (%) of Agaricus bisporus before and after carbonization. F) The contact angle of Agaricus bisporus, carbonized Agaricus bisporus, charcoal, gelatin sponge, and gauze.

Figure 2

The swelling behaviors of different carbonized mushroom aerogels under water and blood. A,B) Photographs of different carbonized mushroom aerogels and gelatin sponges before and after compression, and its shape recovery behaviors after absorbing water or blood. C) The shape recovery time and shape recovery ratio of different carbonized mushroom aerogels and gelatin sponges after absorbing water (n = 3). D) The shape recovery time and shape recovery ratio of different carbonized mushroom aerogels and gelatin sponges after absorbing blood (n = 3). E) The water swelling capacity and swelling speed of different mushroom aerogels and positive controls, including charcoal, gelatin sponge, and Gauze. F) The blood swelling capacity and swelling speed of different mushroom aerogels and positive controls, including charcoal, gelatin sponge, and Gauze. *p < 0.05, ***p < 0.001, ****p < 0.0001.

Figure 3

Mechanical performances of the different carbonized mushroom aerogels. A) Cyclic compression test of the carbonized mushroom aerogels that generated under 160 °C for 12 h, 180 °C for 10 h, and 200 °C for 8 h in 70% compressive strain (20 cycles) respectively. B) The compressive strain–stress curves of the gelatin sponge, and aerogels carbonized under 160 °C for 8, 10, and 12 h respectively. C) The compressive strain–stress curves of the gelatin sponge, and aerogels carbonized for 10 h under 160 °C, 180 °C, and 200 °C respectively. D) The compressive strain–stress curves of the dry and wet aerogels that carbonized under 160 °C for 10 h in 70% compressive strain. E) The tensile strain–stress curves of the gelatin sponge, and aerogels carbonized under 160 °C for 8, 10, and 12 h respectively. F) The tensile strain–stress curves of the gelatin sponge, and aerogels carbonized for 10 h under 160 °C, 180 °C, and 200 °C respectively. G) The tensile strain–stress curves of dry and wet aerogels that carbonized under 160 °C for 10 h in 30% tensile strain.

Figure 4

Biocompatibility evaluation of the carbonized mushroom aerogels in vitro. A,B) The proliferation and cell viability of L929 cells that co‐cultured with extract medium of different carbonized mushroom aerogels for 1, 2, and 3 days, respectively. C) The photographs of hemolysis assay of the charcoal, gelatin sponge, and carbonized mushroom aerogels with different concentrations, ranging from 2.5 to 1.25, and 0.625 mg mL⁻¹. D) The quantification of the hemolysis assay. CT: connective tissues. CMA: carbonized mushroom aerogels.

Figure 5

Hemostatic performances of the different carbonized mushroom aerogels in vitro. A) Photographs of hemoglobin binding capacity of the carbonized mushroom aerogels, gelatin sponge, and Gauze at different detecting times. B) The blood clotting index (BCI (%)) of the carbonized mushroom aerogels, gelatin, and Gauze at different detecting times. C) The whole‐blood clotting time of the carbonized mushroom aerogels, gelatin sponge, and Gauze. The blank group was not exposed to hemostatic materials (n = 3). D) The percentage of adhered RBCs on the surface of carbonized mushroom aerogels, gelatin sponge, and Gauze (n = 3). E) SEM images showing the interfacial reaction on the surface of carbonized mushroom aerogels, gelatin sponge, and Gauze after immersing into whole blood for 1 min. *p < 0.05, **p < 0.01, ****p < 0.0001.

Figure 6

The adhesion and activation of platelets. A) SEM images showing the adhered and activated platelets (shaped changed platelets) on the surface of carbonized mushroom aerogels, gelatin sponge, and Gauze. B) Percentage of adhered platelets on the surface of carbonized mushroom aerogels, and gelatin sponge (n = 3). C) Zeta potential of the carbonized mushroom aerogels, and gelatin sponge (n = 3). D,E) Prothrombin time (PT) and activated partial thromboplastin time (APTT) analysis of the carbonized mushroom aerogels, and gelatin sponge (n = 3). *p < 0.05, **p < 0.01, ****p < 0.0001.

Figure 7

Hemostatic performance of the carbonized mushroom aerogels on the management of rat tail‐amputation and liver‐perforation hemorrhage model. A) Photographs showing the hemostatic ability of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat tail amputation model. The tails without any treatment are set as the blank control group. B) The total blood loss of rats treated with different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat‐tail amputation model (n = 3). C) Hemostatic time of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat tail‐amputation hemorrhage model (n = 3). D) Photographs showing the hemostatic ability of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat liver‐perforation hemorrhage model, the perforated livers without any treatment were set as blank control group. E) The total blood loss of rats treated with different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat liver‐perforation hemorrhage model. F) Hemostatic time of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat liver‐perforation hemorrhage model. *p < 0.05, **p < 0.01, ****p < 0.0001.

Figure 8

Hemostatic performance of the carbonized mushroom aerogels on the management of rabbit cardiac perforation and rat hepatectomized hemorrhage model. A) Photographs showing the hemostatic ability of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rabbit heart uncontrolled hemorrhage model. The heart without any treatment are set as the blank control group. B) The total blood loss of rabbit treated with different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rabbit heart uncontrolled hemorrhage model. C) Hemostatic time of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rabbit heart uncontrolled hemorrhage model. D) Photographs showing the hemostatic ability of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat hepatectomized hemorrhage model. The livers without any treatment were set as blank control group. E) Total blood loss of rat treated with different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat hepatectomized hemorrhage model. F) Hemostatic time of different carbonized mushroom aerogels, Gauze, and gelatin sponge in the rat hepatectomized hemorrhage model. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Figure 9

Histological observations of the injured site after achieving hemostasis. Histological observations of the interfaces between hemostatic materials and A) liver and B) heart tissues after applying them to stop bleeding for 7 days and 14 days. The black dash line refers to the boundary of the injured tissues and the hemostatic materials. CMA: carbonized mushroom aerogels.